Highlighted Paper selected by Editor-in-Chief : Histochemical Imaging of Alkaline Phosphatase Using a Novel Fluorescent Substrate
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- Takahashi Tadanobu
- Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka
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- Otsubo Tadamune
- Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University
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- Ikeda Kiyoshi
- Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University
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- Minami Akira
- Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka
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- Suzuki Takashi
- Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka
書誌事項
- タイトル別名
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- Histochemical Imaging of Alkaline Phosphatase Using a Novel Fluorescent Substrate
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Histochemical visualization of phosphatase is exclusively required for Western immunoblotting and antigen-positive cell staining using an alkaline phosphatase (AP)-labeled secondary antibody. This detection has been performed by several reagents including 5-bromo-4-chloro-3-indolyl-phosphate (X-Phos), nitro blue tetrazolium (NBT), 3-(2′-spiroadamantane)-4-methoxy-4-(3″-phosphoryloxy)phenyl-1,2-dioxetane and 2-(5′-chloro-2′-phosphoryloxyphenyl)-6-chloro-4-[3H]-quinazolinone (ELF® 97 Phosphate). We previously reported that 2-(benzothiazol-2-yl)-4-bromophenol bonded with N-acetylneuraminic acid (BTP3-Neu5Ac), enabled fluorescent histochemical visualization of sialidase activity. 2-(Benzothiazol-2-yl)-4-bromophenol (BTP3), which is formed from BTP3-Neu5Ac by sialidase reaction, is a crystalline, insoluble and stable fluorogenic compound, deposited at the site of enzyme activity. We developed a BTP3 phosphate ester (BTP3-Phos) for the purpose of fluorescent histochemical visualization of phosphatase activity. BTP3-Phos emitted fluorescence in a manner dependent on the concentration of the AP-labeled antibody. BTP3-Phos also enabled fluorescent histochemical visualization of AP-blotted dots in a manner dependent on the concentration of the AP-labeled antibody. The detection sensitivity of BTP3-Phos was estimated to be greater than that of the conventional method using X-Phos and NBT. Influenza A virus-infected cells were fixed and reacted with anti-influenza A virus antibodies and incubated continuously with an AP-labeled secondary antibody. BTP3-Phos stained the infected cells with distinct green fluorescence. These results indicate that BTP3-Phos can enable fluorescent immunohistochemical staining analysis using an AP-labeled antibody. BTP3-Phos would be beneficial for histochemical staining of AP activity, and may be applicable for multi-color staining or a cell sorter.
収録刊行物
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- Biological & Pharmaceutical Bulletin
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Biological & Pharmaceutical Bulletin 37 (10), 1668-1673, 2014
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390282679609275008
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- NII論文ID
- 130004677552
- 40020224783
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- NII書誌ID
- AA10885497
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- COI
- 1:STN:280:DC%2BC2M%2FgtlOrtg%3D%3D
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- ISSN
- 13475215
- 09186158
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- NDL書誌ID
- 025838962
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- PubMed
- 25109307
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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- PubMed
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